First thermostable CLIP-tag by rational design applied to an archaeal O⁶ -alkyl-guanine-DNA-alkyl-transferase.

ABSTRACT: Self-labelling protein tags (SLPs) are resourceful tools that revolutionized sensor imaging, having the versatile ability of being genetically fused with any protein of interest and undergoing activation with alternative probes specifically designed for each variant (namely, SNAP-tag, CLIP-tag and Halo-tag). Commercially available SLPs are highly useful in studying molecular aspects of mesophilic organisms, while they fail in characterizing model organisms that thrive in harsh conditions. By applying an integrated computational and structural approach, we designed a engineered variant of the alkylguanine-DNA-alkyl-transferase (OGT) from the hyper-thermophilic archaeon Saccharolobus solfataricus (SsOGT), with no DNA-binding activity, able to covalently rea

SUBMITTER: Merlo R 

PROVIDER: S-EPMC9519396 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature